Kulle Alexandra E, Reinehr Thomas, Simic-Schleicher Gunter, Hornig Nadine C, Holterhus Paul-Martin
Department of Pediatrics, Division of Pediatric Endocrinology and Diabetes, Christian-Albrechts-University Kiel and University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Department of Pediatric Endocrinology, Diabetes and Nutrition Medicine, Vestische Hospital for Children and Adolescents Datteln, University of Witten/Herdecke, Datteln, Germany; and.
J Clin Endocrinol Metab. 2017 Jan 1;102(1):232-241. doi: 10.1210/jc.2016-2849.
Dehydroepiandrosterone sulfate (DHEAS) and 17-hydroxypregnenolone (17OHPreg) are important for understanding the Δ5 pathway (e.g., in adrenarche and obesity). Although mass spectrometry has become the state-of-the-art method for quantifying steroids, there are few comprehensive age-, sex-, and pubertal stage-specific reference ranges for children.
To develop a sensitive and reliable ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous quantification of DHEAS and 17OHPreg and to establish entire age-, sex- and pubertal stage-specific reference ranges in children.
A total of 684 children, 453 (243 female, 210 male) with normal body mass index (BMI; <90th) and 231 (132 female, 99 male) obese subjects (>97th), were categorized into 11 age groups, and age- and Tanner stage (PH)-specific reference ranges were determined.
The limit of detection was 0.05 nmol/L for 17OHPreg and 0.5 nmol/L for DHEAS. Levels of both steroids declined after the neonatal period. Comparisons with RIA assays (Siemens, Munich, Germany) (DHEAS) and an in-house kit (17OHPreg) revealed 0.95 and 0.93, respectively, as coefficients of determination. Although DHEAS-generally higher in boys-increased continuously starting at 3 to 6 years, 17OHPreg remained largely constant. In obese patients, both were significantly elevated, also in part after alignment to Tanner stages (PH).
UPLC-MS/MS is sensitive and reliable for quantifying DHEAS and 17OHPreg. Our data support differential maturation of CYP17 during adrenarche with successively increasing 17,20-lyase activity but largely constant 17α-hydroxylation activity. Endocrine interpretation of 17OHPreg and DHEAS must consider differential patterns for age, sex, pubertal stage, and BMI.
硫酸脱氢表雄酮（DHEAS）和17-羟孕烯醇酮（17OHPreg）对于理解Δ5途径（例如在肾上腺初现和肥胖中）很重要。尽管质谱分析法已成为类固醇定量的先进方法，但针对儿童的综合年龄、性别和青春期阶段特异性参考范围却很少。
开发一种灵敏且可靠的超高效液相色谱串联质谱法（UPLC-MS/MS），用于同时定量DHEAS和17OHPreg，并建立儿童全年龄、性别和青春期阶段特异性参考范围。
总共684名儿童，其中453名（243名女性，210名男性）体重指数正常（BMI；<第90百分位数），231名（132名女性，99名男性）肥胖受试者（>第97百分位数），被分为11个年龄组，并确定了年龄和坦纳分期（PH）特异性参考范围。
17OHPreg的检测限为0.05 nmol/L，DHEAS的检测限为0.5 nmol/L。两种类固醇的水平在新生儿期后均下降。与放射免疫分析（RIA）检测（德国慕尼黑西门子公司）（DHEAS）和内部试剂盒（17OHPreg）的比较显示，决定系数分别为0.95和0.93。尽管DHEAS通常在男孩中较高，从3至6岁开始持续升高，但17OHPreg基本保持恒定。在肥胖患者中，两者均显著升高，部分也是在根据坦纳分期（PH）进行调整之后。
UPLC-MS/MS对于定量DHEAS和17OHPreg灵敏且可靠。我们的数据支持肾上腺初现期间CYP17的差异成熟，17,20-裂解酶活性逐渐增加，但17α-羟化活性基本恒定。对17OHPreg和DHEAS的内分泌学解释必须考虑年龄、性别、青春期阶段和BMI的差异模式。
